Detection of the nuclear poly(ADP-ribose)-metabolizing enzymes and activities in response to DNA damage.
Poly(ADP-ribosyl)ation is a posttranslational modification of proteins in higher eukaryotes mediated by poly(ADP-ribose) polymerases (PARPs) that is involved in many physiological processes such as DNA repair, transcription, cell division, and cell death. Biochemical studies together with PARP-1- or PARP-2-deficient cellular and animal models have revealed the redundant but also complementary functions of the two enzymes in the surveillance and maintenance of genome integrity. Poly(ADP-ribose) is degraded by the endo- and exo-glycosidase activities of poly(ADP-ribose) glycohydrolase (PARG). In this chapter, biochemical and immunofluorescence methods are described for detecting and assaying PARPs and PARG.
Amé
Jean-Christophe
JC
Département Intégrité du Génome, UMR 7175-LCI du CNRS, Ecole Supérieure de Biotechnologie de Strasbourg, Illkirch, France.
Hakmé
Antoinette
A
Quenet
Delphine
D
Fouquerel
Elise
E
Dantzer
Françoise
F
Schreiber
Valérie
V
D000818:Animals; D015153:Blotting, Western; D004249:DNA Damage / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism
D011064:Poly Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases; D006026:Glycoside Hydrolases; C022407:poly ADP-ribose glycohydrolase